In this manuscript, Mikako et al. showed promising in vitro results of the effects of PBK inhibition in cancer cells, and explored the molecular mechanism by which this is achieved. The manuscript is well written and the data provided looks very interesting.

I have one major concern. Why for some experiments, for example the ones shown in figures 1A, 3,4, there is data provided from all 4 cell lines, whit consistent results within each other. But then for other experiments, the data shown is not from all 4 cell lines, just from some of them. For example in figure 1B, no data is provided for the cell line OSC-19, in figure 2A, cell lines SAS and OSC-19 are missing, figure 2C, cell line SAS is missing, and figure 5, cell lines HSC-3 and OSC-19 are missing. Please explain this rationale.

Minor comments:
Introduction:

The HPV paragraph in the introduction feels a little bit out of context and incomplete. It explains how E7 protein could inactivate the retinoblastoma protein, but no mention on how E6 could affect p53.

MM

Why are the concentrations of OTS514 so different between the different experiments? The reported IC50 doses obtained from the proliferation assay are much higher than the ones used in the other experiments. Please explain this.

The authors have clarified all my concerns

The authors have addressed all my suggestions

The authors have clarified all my concerns